Konsorsium Riset Vaksin TB Center for Biomedical and Basic Technology of Health National Institute of Health Research and Development Solo, 26-27 Oktober 2017 Seminar Nasional Riset TB Vaksin TB Protein Sub-Unit sebagai Booster Vaksinasi BCG di Indonesia
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Konsorsium Riset Vaksin TB
Center for Biomedical and Basic Technology of Health
National Institute of Health Research and Development
Solo, 26-27 Oktober 2017
Seminar Nasional Riset TB
Vaksin TB Protein Sub-Unit
sebagai Booster Vaksinasi BCG
di Indonesia
Outline
• Vaksin BCG
• Pilihan protein sub-unit sebagai kandidat
booster vaksin BCG
• Capaian Riset Vaksin TB
• Kolabrasi – produk tambahan
Konsorsium Riset Vaksin TB
Anggota
13 Institusi
PT Bio Farma PT Pakar Biomedika
Indonesia
Riset Prioritas Nasional - BAPPENAS
Koordinator
PBTDK
Riset Dasar • M.tb “dorman”
• Model Granuloma in vitro
• Interaksi human - patogen
Riset Produk • Vaksin
• Kit Diagnostik
• Produk Biosimilar / Model
2012 - 2020
Vaksin BCG Pasteur
Pre-infection
Post-infection
Post-infection/
therapeutic
TB Vaccine rethinking
• The life cycle of TB infection/disease and its impact on
vaccination strategies
• Host–pathogen interaction in various stages of TB infection
– Cellular dynamics in different stages of TB infection
– Antigen expression in different stages of infection
• Immunological memory and TB protection
• Vaccine strategies against TB
– Pre-exposure vaccines
• Viable mycobacteria are designed to replace BCG as prime vaccines
• Subunit vaccines comprise MTB protein antigens expressed in viral
vectors or delivered in adjuvant and are designed as BCG boosters
– Post-exposure vaccines target adolescents and adults with LTBI
Pendekatan Pengembangan Vaksin Sub-Unit
• Early secreted protein
– RD1 deletion (ESAT6, CFP10 dll)
• PE/PPE family
– Highly antigenic – function?
– 10% of genome
• Gap analysis of BCG Vaccine Pasteur vs Tokyo
sequences (results of WGS approach)
• Proteomics analysis of “granuloma in vitro” medels
Output I - Riset TB
• Protein Sub-Unit/Antigen Kandidat Vaksin
– Ag85B-Rv2660c
– Ag85B
– Esat6-Cfp10
– Esat6-Mtb32c-Cfp10
– RpfB & RpfD
– PPE41 & PPE17
– PE-PGRS 14, 24, 32, 35, 45
– LipY
– ManLam
Diserahkan ke PT Bio Farma
• Protein/Klon
• MTA
• Seed history
Hasil Purifikasi
Pelaksana: PBTDK, ITB, UI, UH, UM, UGM, LIPI
PE/PPE Protein
• PPE41
– Antigenik, disekresi bersama sistem Esx
sebagai kandidat vaksin TB. Optimasi dg
PPE25 (esx system) soluble protein
– Dormancy
• PPE17 - Diagnostic tools
Whole Genome Sequencing Vaksin BCG Pasteur
BCG Pasteur Biofarma sesuai
Parent seed
Terdapat 5 gen yang berbeda
antara BCG Pasteur dengan
BCG Tokyo
Pemilihan antigen kandidat
vaksin TB sesuai dengan BCG
Tokyo yang lebih efektif
• Rv0490&Rv0491,
• Rv1189,
• Rv1441&Rv1325
Model “granuloma in vitro”
• Interaksi human-pathogen
– Model granuloma in vitro
• PBMC - M.tb
• Ditambah matriks extraseluler
• Indikator Model Granuloma
– CD68 (macrophage) & CD3 & CD63 (sel T)
– Mikroskopik
• Analisis Proteomiks (LC MS/MS)
– Hari 1 dan hari 7, dengan dan tanpa M.tb
• M.tuberculosis dorman in vitro
– Kultur – deplesi O2, minimal nutrition
Penilaian M.tb dorman in vitro pada
pembentukan granuloma
• Agregasi sel – Granuloma like:
– Dibuat dari PBMC ditambahkan M.tb H37Rv
inkubasi selama 3 hari (waktu penilaian)
• Fenotipik: Gambaran terbentuknya granuloma dengan indeks
kepadatan sel (index score) dibuat kriteria nya menggunakan
mikroskop inverted. Morfologi MO dan limfosit dengan HE
• Reaktivasi dengan menilai faktor virulensi bakteri berdasarkan enzim metabolik: ICL (isocitrate lyase) (Rv0467) – upregulasi pada jalur metabolisme lemak merupakan marker dormancy
– Dorman • Pembentukan granuloma: TNF dan IFN (minimal)
Indikator
Biomarkers of protection against TB, following
BCG vaccination
IL-2
IL-17
IFN-
TNF
Classical Th1 responses do not
associate with risk of TB disease, after
BCG
*BCG given at birth. Infants followed for 2 years to assess
protection; Results shown from whole blood incubation